Diagnostic and prognostic value of peripheral blood leucocyte ratios in sick cats

The objective of this study was to assess the diagnostic and prognostic utility of feline neutrophil-to-lymphocyte (NLR) and lymphocyte-to-monocyte ratios (LMR) in a variety of underlying diseases. Five-year medical records from cats presenting to the internal medicine unit of a veterinary teaching hospital were retrospectively reviewed. Cats were considered for inclusion based on complete medical records. ADVIA 120 was used for the complete blood counts; the NLR and LMR were calculated by dividing the absolute numbers of the respective leucocytes. Two hundred and nineteen sick and 20 healthy cats were included in the study. The median NLR and LMR were significantly (P < 0.05) elevated and decreased, respectively, in cats with infectious, neoplastic and chronic kidney diseases compared to controls. Additionally, cats with neoplasia had significantly higher median NLR compared to cats with urinary tract and gastrointestinal diseases. Non-survivors had significantly higher NLR and lower LMR compared to survivors. Both ratios had suboptimal prognostic performance for the outcome of sick cats (NLR sensitivity: 37.9%, specificity: 86.4%; LMR sensitivity: 69.0%, specificity: 61.0%). Many different disease categories were associated with increased NLR and decreased LMR compared to controls, but the overall prognostic performance of the two leucocyte ratios was suboptimal.

Peripheral blood leucocyte telomere length is associated with progression of interstitial lung disease in systemic sclerosis

BackgroundPeripheral blood leucocyte telomere length (PBL-TL) is associated with outcomes in patients with idiopathic pulmonary fibrosis. Whether PBL-TL is associated with progression of systemic sclerosis-associated interstitial lung disease (SSc-ILD) is unknown.MethodsA retrospective observational cohort study was performed using prospectively collected data from 213 patients with SSc followed at the University of California San Francisco (UCSF) Scleroderma Center. PBL-TL was measured by quantitative PCR of DNA isolated from peripheral blood. Associations between PBL-TL and pulmonary function test trends in patients with SSc-ILD were assessed by longitudinal analysis using Generalised Linear Mixed Models. Findings were validated in a cohort of 61 patients with SSc-ILD enrolled in the Stanford University Scleroderma Center database.ResultsPatients with UCSF SSc with ILD were found to have shorter PBL-TL compared with those without ILD (6554±671 base pairs (bp) vs 6782±698 bp, p=0.01). Shorter PBL-TL was associated with the presence of ILD (adjusted OR 2.1 per 1000 bp TL decrease, 95% CI [1.25 to 3.70], p=0.006). PBL-TL was shorter in patients with SSc-ILD lacking SSc-specific autoantibodies compared with seropositive subjects (6237±647 bp vs 6651±653 bp, p=0.004). Shorter PBL-TL was associated with increased risk for lung function deterioration with an average of 67 mL greater loss in per year for every 1000 bp decrease in PBL-TL in the combined SSc-ILD cohorts (longitudinal analysis, adjusted model: 95% CI −104 mL to −33 mL, p<0.001).ConclusionsThese findings suggest that telomere dysfunction may be associated with SSc-ILD progression and that PBL-TL measurement may be useful for stratifying risk for SSc-ILD progression.

Artificial intelligence and leukocyte epigenomics: Evaluation and prediction of late-onset Alzheimer’s disease

We evaluated the utility of leucocyte epigenomic-biomarkers for Alzheimer’s Disease (AD) detection and elucidates its molecular pathogeneses. Genome-wide DNA methylation analysis was performed using the Infinium MethylationEPIC BeadChip array in 24 late-onset AD (LOAD) and 24 cognitively healthy subjects. Data were analyzed using six Artificial Intelligence (AI) methodologies including Deep Learning (DL) followed by Ingenuity Pathway Analysis (IPA) was used for AD prediction. We identified 152 significantly (FDR p<0.05) differentially methylated intragenic CpGs in 171 distinct genes in AD patients compared to controls. All AI platforms accurately predicted AD with AUCs ≥0.93 using 283,143 intragenic and 244,246 intergenic/extragenic CpGs. DL had an AUC = 0.99 using intragenic CpGs, with both sensitivity and specificity being 97%. High AD prediction was also achieved using intergenic/extragenic CpG sites (DL significance value being AUC = 0.99 with 97% sensitivity and specificity). Epigenetically altered genes included CR1L & CTSV (abnormal morphology of cerebral cortex), S1PR1 (CNS inflammation), and LTB4R (inflammatory response). These genes have been previously linked with AD and dementia. The differentially methylated genes CTSV & PRMT5 (ventricular hypertrophy and dilation) are linked to cardiovascular disease and of interest given the known association between impaired cerebral blood flow, cardiovascular disease, and AD. We report a novel, minimally invasive approach using peripheral blood leucocyte epigenomics, and AI analysis to detect AD and elucidate its pathogenesis.

Standard protocols for immune profiling of peripheral blood leucocyte subsets by flow cytometry using DuraClone IM reagents

This document describes standard protocols used by the Hutchinson group for profiling human peripheral blood leucocytes by flow cytometry analyses using DuraClone IM reagents.

The morphological response of the heart and spleen following acute myocardial infarction-induced sterile inflammation: a clinicopathological study

Introduction: as an outcome of sterile inflammation-inducing acute ischemic processes, many splenic cells enter the circulatory system and migrate into the lesion, defending tissues against the spread of ischemia or enhancing necrosis.Objective: investigating the therapeutic effect of splenic morphological response to sterile inflammation-inducing myocardial infarction.Material and method: We examined the weight and structure of the heart and spleen of 106 patients deceased due to acute myocardial infarction. These data were correlated with demographic (personal) and epidemiological data, and disease history. After morphological investigation of archived myocardial and splenic tissue samples, the acute ischemia-induced structural alterations of splenic samples were quantified using a digital morpho-metric method. Results were evaluated in comparison to the myocardial ischemia coefficient. Changes in distribution of ischemia-induced cell types were characterized by defining the immunological phenotypes of macrophages (M1 vs. M2). Spleen samples from patients without history of ischemia were used as controls.Results: The modification of the spleen weight was associated with an increase in peripheral blood leucocyte levels. Our morphological analysis proved a positive correlation between the ischemia coefficient and the decrease of spleen weight. Structural analysis of splenic tissue revealed the collapse of red pulp sinusoids, a significant size decrease of the white pulp marginal zone (p<0.05), and depleted follicles with irregular margins without any distinct germinative centers. Concurrently, with the proliferation of granulocytes, the increase of M1 macrophages was observed in the myocardium, and a higher M1/M2 ratio was detected in the marginal zone of splenic follicles.Conclusion: On the background of acute ischemia, time critically determines the dynamic structural changes of the spleen. Along with reducing the marginal zone, immunomodulation targeting its cellular composition will be a putative therapeutic approach in the future.

Comparison of leucocyte profiles between healthy children and those with asymptomatic and symptomatic Plasmodium falciparum infections

Background The immune mechanisms that determine whether a Plasmodium falciparum infection would be symptomatic or asymptomatic are not fully understood. Several studies have been carried out to characterize the associations between disease outcomes and leucocyte numbers. However, the majority of these studies have been conducted in adults with acute uncomplicated malaria, despite children being the most vulnerable group. Methods Peripheral blood leucocyte subpopulations were characterized in children with acute uncomplicated (symptomatic; n = 25) or asymptomatic (n = 67) P. falciparum malaria, as well as malaria-free (uninfected) children (n = 16) from Obom, a sub-district of Accra, Ghana. Leucocyte subpopulations were enumerated by flow cytometry and correlated with two measures of parasite load: (a) plasma levels of P. falciparum histidine-rich protein 2 (PfHRP2) as a proxy for parasite biomass and (b) peripheral blood parasite densities determined by microscopy. Results In children with symptomatic P. falciparum infections, the proportions and absolute cell counts of total (CD3 +) T cells, CD4 + T cells, CD8 + T cells, CD19 + B cells and CD11c + dendritic cells (DCs) were significantly lower as compared to asymptomatic P. falciparum-infected and uninfected children. Notably, CD15 + neutrophil proportions and cell counts were significantly increased in symptomatic children. There was no significant difference in the proportions and absolute counts of CD14 + monocytes amongst the three study groups. As expected, measures of parasite load were significantly higher in symptomatic cases. Remarkably, PfHRP2 levels and parasite densities negatively correlated with both the proportions and absolute numbers of peripheral leucocyte subsets: CD3 + T, CD4 + T, CD8 + T, CD19 + B, CD56 + NK, γδ + T and CD11c + cells. In contrast, both PfHRP2 levels and parasite densities positively correlated with the proportions and absolute numbers of CD15 + cells. Conclusions Symptomatic P. falciparum infection is correlated with an increase in the levels of peripheral blood neutrophils, indicating a role for this cell type in disease pathogenesis. Parasite load is a key determinant of peripheral cell numbers during malaria infections.

Exploration of prognostic factors for critical COVID-19 patients：a nomogram analysis

To discuss influencing factors on critical COVID-19 patient’s prognosis, construct a basic model and predict their mortality risks. Retrospectively analyzed the general condition and respective laboratory biomarkers of critical patients with duration≥24 h from Feb. 10th, 2020 to Mar. 30th, 2020 to separate them into a survival group and death group based on their clinical features. Multiple logistic regression analysis was performed to assess risk factors for critical COVID-19 patient’s and a nomogram was constructed based on screened risk factors. A receiver operating curve (ROC) was created to evaluate the accuracy of the nomogram. Multi-factor Logistic recovery analysis results show: Age, Peripheral blood leucocyte count,Lymphocyte percentage, Thrombocyte count and Hyper C-reactive protein are single danger factors of critical COVID-19 patient’s mortality risk (p＜0.05). ROC curve indicates Nomogram predictive model AUC is 0.958 (95%CI: 0.923-0.993), which has high predictive value. Findings from this study suggest advanced age, high peripheral blood leucocyte count, high hyper C-reactive protein, low lymphocyte percentage and low thrombocyte count are risk factors of critical COVID-19 patient’s death.The Nomogram model is helpful for timely intervention to reduce the incidence of critical COVID-19 patients.

SAT-197 Unilateral Adrenalectomy Can Induce Control in ARMC5 Mutations Patients

Primary macronodular hyperplasia (PMAH) is a rare cause of endogenous Cushing’s syndrome characterized by functioning adrenal macronodules and variable cortisol secretion. ARMC5 is the most frequent gene responsible for PMAH. Genetic mutations including inactivating germline mutations in armadillo repeat-containing 5 (ARMC5) gene have been identified. The occurrence of several other non adrenal tumors (meningioma, breast, colon, thyroid and parathyroid) has also been associated with PBMAH, suggesting a possible role of ARMC5 for the development of other neoplasias. The best treatment to this condition is not established. Clinical case: 64-year-old man, referred due to bilateral adrenal incidentalomas on a CT scan with characteristics of adenoma/hyperplasia. He had type-2 diabetes, hypertension and dyslipidemia for about 4 years. He was being treated with 4 different anti-hypertensive medications, gliclazide and simvastatin. Physical examination revealed thin and dry skin, obesity with centripetal fat distribution, multiple ecchymosis and facial erythrosis. He had a son and a daughter. No known familial relevant diseases. His laboratory workup revealed ACTH-independent hypercortisolism: failure to suppress on the overnight dexametasone suppression test - cortisol 27.6ug/dL- and on the low-dose dexamethasone test- cortisol 24ug/dL- associated to morning ACTH &lt;1ng/L. He underwent stimulation tests with tetracosactide, LHRH, TRH, vasopressin, metoclopramide, glucagon anddeambulation test, which were overall negative. It was decided to proceed to bilateral adrenalectomy. Due to surgical complications, only right adenalectomy was performed. Histology revealed adrenal nodular hyperplasia. Molecular study in DNA extracted from peripheral blood leucocyte and in the adrenal gland revealed the presence in heterozygosity of the pathogenic mutation c.1379T&gt;C in the ARMC5 gene. The patient underwent a cerebral CT scan which showed a meningioma in the posterior left temporal convexity. Genetic testing was also performed on the daughter and son of the patient, which revealed the same mutation. They were also tested for hypercortisolism and underwent adrenal and cerebral CT scan, which showed no abnormality. The adrenal CT scan of the daughter showed enlargement of the left adrenal gland, with aspects suggestive of nodular hyperplasia. Her blood tests revealed no sign of hypercortisolism - overnight dexametasone test- cortisol 1.0 ug/dL, morning ACTH 19.1 ng/L. The patient is currently with cortisol hypersecretion controlled. Conclusion: All patients with PMAH should be tested for ARMC5 mutations and if they are found, family screening is mandatory in this autosomal dominant disorder. Unilateral adrenalectomy could control the cortisol hypersecretion, however some of these patients can have subclinical Cushing Syndrome.